Rapid fluidic exchange microsystem for recording of fast ion channel kinetics in Xenopus oocytes.
We present a new lab-on-a-chip system for electrophysiological measurements on Xenopus oocytes. Xenopus oocytes are widely used host cells in the field of pharmacological studies and drug development. We developed a novel non-invasive technique using immobilized non-devitellinized cells that replaces the traditional "two-electrode voltage-clamp" (TEVC) method. In particular, rapid fluidic exchange was implemented on-chip to allow recording of fast kinetic events of exogenous ion channels expressed in the cell membrane. Reducing fluidic exchange times of extracellular reagent solutions is a great challenge with these large millimetre-sized cells. Fluidic switching is obtained by shifting the laminar flow interface in a perfusion channel under the cell by means of integrated poly-dimethylsiloxane (PDMS) microvalves. Reagent solution exchange times down to 20 ms have been achieved. An on-chip purging system allows to perform complex pharmacological protocols, making the system suitable for screening of ion channel ligand libraries. The performance of the integrated rapid fluidic exchange system was demonstrated by investigating the self-inhibition of human epithelial sodium channels (ENaC). Our results show that the response time of this ion channel to a specific reactant is about an order of magnitude faster than could be estimated with the traditional TEVC technique.
Dahan
E
E
Ecole Polytechnique Fédérale de Lausanne, CH-1015 Lausanne, Switzerland.
Bize
V
V
Lehnert
T
T
Horisberger
J-D
JD
Gijs
M A M
MA
D000818:Animals; D004563:Electrochemistry; D055724:Electrophysiological Phenomena / Q000187:drug effects; D004847:Epithelial Cells / Q000187:drug effects / Q000378:metabolism; D053503:Epithelial Sodium Channels / Q000235:genetics / Q000378:metabolism*; D005260:Female; D006801:Humans; D007700:Kinetics; D046210:Microfluidic Analytical Techniques / Q000379:methods*; D009865:Oocytes / Q000166:cytology / Q000378:metabolism*; D018408:Patch-Clamp Techniques; D010477:Perfusion; D012964:Sodium / Q000378:metabolism / Q000494:pharmacology; D015222:Sodium Channels / Q000378:metabolism; D013997:Time Factors; D014981:Xenopus
D053503:Epithelial Sodium Channels; D015222:Sodium Channels; D012964:Sodium